Determination of kinetic parameters for 3α-hydroxysteroid dehydrogenase using the five major bile acids and their conjugates as substrates and correlation with their structure and solubility

The Analyst ◽  
1987 ◽  
Vol 112 (6) ◽  
pp. 753-755 ◽  
Author(s):  
Evriklia S. Lianidou ◽  
Panayiolis A. Siskos
Keyword(s):  
Kinetic Parameters ◽  
Bile Acids ◽  
Hydroxysteroid Dehydrogenase

Effect of protein on the determination of total bile acids in serum.

1983 ◽  
Vol 29 (1) ◽  
pp. 171-175 ◽  
Author(s):  
N Q Hanson ◽  
E F Freier
Keyword(s):  
Bile Acid ◽  
Bile Acids ◽  
Hydroxysteroid Dehydrogenase ◽  
Inhibitory Effect ◽  
Normal Serum ◽  
Total Serum ◽  
Enzyme Preparations ◽  
Analytical Recovery ◽  
Fluorescence Light

Abstract We measured total serum bile acids on a fluorescence-light-scattering micro centrifugal analyzer by the direct enzymatic method with 3 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.50) and with resazurin as a fluorogenic electron acceptor. We found that serum protein has an inhibitory effect on the measurement of bile acids, but this effect was eliminated by adding bovine serum albumin to the reaction mixture in a final protein concentration (12.2 g/L) that was high compared with that contributed by a normal serum specimen. The assay is a sensitive method that reaches equilibrium in 5 min. The method is microscale (5 microL of sample, 150 microL of working reagent), is easy to perform, and is accurate (analytical recovery = 104.1%) and precise (CV = 11.1 and 5.7% on specimens with bile acid concentrations of 7.6 and 35.4 mumol/L, respectively). Normal values are 1-12 and less than 9 mumol/L on nonfasting and fasting individuals, respectively. Pure 3 alpha-hydroxysteroid dehydrogenase must be used: we found several enzyme preparations that gave falsely high values for bile acid.

A Direct Enzymic Assay for 7α-Hydroxy Bile Acids and Their Conjugates

1973 ◽  
Vol 44 (1) ◽  
pp. 95-98 ◽  
Author(s):  
G. A. D. Haslewood ◽  
G. M. Murphy ◽  
Judith M. Richardson
Keyword(s):  
Thin Layer ◽  
Bile Acids ◽  
Thin Layer Chromatography ◽  
Biological Fluids ◽  
Hydroxysteroid Dehydrogenase ◽  
Layer Chromatography ◽  
Enzymic Assay ◽  
Bile Acids And Salts ◽  
Secondary Bile Acids

1. A 7α-hydroxysteroid dehydrogenase preparation isolated from a strain of Escherichia coli provides a specific and rapid means for the determination of 7α-hydroxy bile acids and their conjugates. 2. When used in conjunction with 3α-hydroxysteroid dehydrogenase the method enables the concentrations of primary and secondary bile acids and salts in biological fluids to be determined directly without the use of thin-layer chromatography. 3. When used with thin-layer chromatography the method allows the specific quantitative determination of chenodeoxycholic acid or its conjugates in the presence of deoxycholic acid and its derivatives.

An enzymatic cycling method for the determination of serum total bile acids with recombinant 3α-hydroxysteroid dehydrogenase

2004 ◽  
Vol 326 (1) ◽  
pp. 87-92 ◽  
Author(s):  
Guo-Hua Zhang ◽  
Ai-Ri Cong ◽  
Guo-Bing Xu ◽  
Chuan-Bao Li ◽  
Rui-Feng Yang ◽  
...  
Keyword(s):  
Bile Acids ◽  
Hydroxysteroid Dehydrogenase ◽  
Enzymatic Cycling ◽  
Total Bile Acids

Determination of bile acids and their conjugates in serum by HPLC using immobilized 3α-hydroxysteroid dehydrogenase for detection

1989 ◽  
Vol 3 (2) ◽  
pp. 75-78 ◽  
Author(s):  
G. R. Campbell ◽  
G. W. Odling-Smee ◽  
B. J. Rowlands ◽  
G. B. Irvine
Keyword(s):  
Bile Acids ◽  
Hydroxysteroid Dehydrogenase
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